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Differentiation of Intestinal Organoids in Cultrex Synthetic Hydrogel.
Organoids expanded in Cultrex Synthetic Hydrogel exhibit expected cystic morphology, indicative of healthy baseline development. Upon differentiation, the organoids begin to lose their cystic morphology and start forming more specialized, structured tissues. This demonstrates the ability of Cultrex Synthetic Hydrogel to support both healthy initial organoid growth and subsequent differentiation into mature structures.
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Cultrex Synthetic Hydrogel Performance Compared to Basement Membrane Extracts.
Cultrex UltiMatrix BME (Catalog # BME001-05) domes were plated at 10,000 cell per 10 μL hydrogel, and Cultrex Synthetic Hydrogel (Catalog # CSH-RUO) domes were plated at 15,000 cells. After three days in culture, comparable organoid growth is observed.
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Cultrex Synthetic Hydrogel Performance Compared to Competitor Basement Membrane Extracts.
Domes using animal component derived matrix from another supplier and Cultrex Synthetic Hydrogel (Catalog # CSH-RUO) were each plated at 10,000 cell per 10 μL hydrogel. Morphologically similar organoid growth is observed after 4 and 5.5 days, respectively.
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Cultrex Synthetic Hydrogel Outperforms Alternative Synthetic Matrices.
Successful organoid growth observed in Cultrex Synthetic Hydrogel (Catalog # CSH-RUO-01) while organoids encapsulated in a competitor's defined hydrogel showed failed organoid formation and significant cell death by day 4.
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Defined Matrix Supports Organoid Growth.
Adult airway organoids were cultured in Cultrex Synthetic Hydrogel (Catalog # CSH-RUO-01) with Recombinant Human FGF-10 Heat Stable Protein (Catalog # BT-FGF10HS). Scale bar indicates 1 mm.
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ICC Imaging of Enteroids.
Images of human enteroids expanded with Wnt/RSPO2 (Catalog # BT-WRSP2) in Cultrex Synthetic Hydrogel (Catalog # CSH-RUO-01), with quantification of total organoid area on day 4. ICC images of enteroids stained with Villin (Catalog # NBP2-75707), E-Cadherin (Catalog # AF748) and DAPI in Synthetic Hydrogel. Scale bar indicates 50 µm.
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Forskolin Swelling Assay.
Enteroids cultured in Wnt/RSPO2 (Catalog # BT-WRSP2) and Synthetic Hydrogel (Catalog # CSH-RUO-01) were differentiated for 7 days and then exposed to Forskolin (10 µM, Catalog # 1099) or DMSO (control) to assess the function of mature CFTR-expressing secretory cells (e.g. enterocytes). The presence of Forskolin led to organoid swelling whereas control did not change in size. Scale bar indicates 100 µm.
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ICC Imaging of Lung Sections.
Lung organoids grown with FGF-10 HS (Catalog # BT-FGF10HS) in Cultrex Synthetic Hydrogel (Catalog # CSH-RUO-01). Sections of lung organoids (cultured for 25 days) in Synthetic Hydrogel with FGF-10 HS stained for SFTPC, SCGB1A1, FOXJ1 (Catalog # AF3619), and DAPI. Scale bar indicates 50 µm.
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Brightfield Images of Liver Organoids Under Expansion and Differentiation Conditions.
Liver organoids were cultured in Cultrex Synthetic Hydrogel (Catalog # CSH-RUO-01) under expansion (top row, scale bar indicates 500 µm) and differentiation conditions (bottom row, scale bar indicates 100 µm). Expanding liver organoids exhibit typical cystic morphology and can be passaged at least three times in Synthetic Hydrogel (data not shown). Upon differentiation, the organoids transition to a more complex structure, displaying folding and wrinkled morphologies.
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